Control of cell arrangement

ABSTRACT

A device for controlling cell arrangement having a pattern composed of a cell adhesive surface and a cell non-adhesive surface, which has a good selectivity of cell adhesion and which can provide a fine pattern of cells in high resolution by culturing cells in a usual manner. The device is prepared by a process which comprises applying a photosensitive, cell adhesive or cell non-adhesive polymer to a surface having a reverse cell adhesivity property to that of the applied polymer, and patternwise irradiating a light to the photosensitive polymer layer followed by development; a process which comprises patternwise irradiating UV or radiation to a cell non-adhesive surface to produce a cell adhesive functional group such as carboxyl group or amino group in the irradiated portion; or a process which comprises patternwise irradiating UV or radiation to a cell adhesive or non-adhesive surface made of plastics to produce a polymerization initiation site in the irradiated portion, and graft-polymerizing at least one monomer having the reverse cell adhesivity property to the plastic surface to produce a polymer on the irradiated portion of plastic surface.

BACKGROUND OF THE INVENTION

The present invention relates of control of the arrangement of cells,and more particularly to a device for controlling cell arrangement, aprocess for preparing the device and a process for controlling the cellarrangement.

In recent years, with rapid progress of cell technology, technology inmanufacturing an LSI and medical technology, attention is attracted ondevices using cells, such as ultra-small biosensor, switching device,bioreactor, hybrid type artificial organs and neuro-computer, anddevelopment of them has been lively made.

It has been difficult to arrange cells as desired and keeping thefunctions thereof, and the difficulty is one of obstacles to realizationof devices using cells. The cell arrangement control technique forarranging the cells as desired, for instance, in a circuit network wouldbecome a key technology for realizing these devices.

It has been attempted to control cell arrangement by patternwiseapplying fibronectin which is a cell adhesive protein, using an ink jetprinter, and culturing cells on the protein pattern. However, thisprocess is poor in resolution and uniformity and is not suitable forfine processing. Recently, control of direction of neuron cell synapsegrowth is attempted using an artificial uneven surface, but formation ofa desirable arrangement is not achieved.

It is a primary object of the present invention to provide a techniquefor easily controlling the arrangement of cells.

The above and other objects of the present invention will becomeapparent from the description hereinafter.

SUMMARY OF THE INVENTION

It has now been found that arrangement of cells can be easily controlledby culturing cells on a material having a pattern composed of a celladhesive surface and a cell non-adhesive surface, whereby a desirablecell pattern can be easily obtained, and that a device having a patternfor cell arrangement composed of the cell adhesive surface and the cellnon-adhesive surface, which is suitable for use in formation of a cellpattern, can be easily prepared through specific steps.

In an aspect of the present invention, there is provided a device forcontrolling cell arrangement having a pattern composed of a celladhesive surface and a cell non-adhesive surface. When cells arecultured on the device, cells grow only on the cell adhesive surface,thus a pattern of cells is obtained.

The patterned cell adhesive surface can be easily obtained by utilizinga photosensitive polymer having a cell adhesivity or a cellnon-adhesivity. Accordingly, in another aspect of the present invention,there is provided a process for preparing a cell arrangement controldevice which comprises applying, onto a surface having either a celladhesivity or a cell non-adhesivity, a photosensitive polymer having thereverse property to said surface with respect to the cell adhesivitycharacteristic, irradiating the resulting photosensitive polymer layerpatternwise with a light, developing the irradiated layer to form apattern in the polymer layer so that the cell adhesive or non-adhesivesurface appears in the non-irradiated portion.

Patternwise irradiation of a cell adhesive or non-adhesive surface withultraviolet rays or radiation is also effective for forming thepatterned cell adhesive surface.

Accordingly, in another aspect of the present invention, the cellarrangement control device is prepared by patternwise irradiating a cellnon-adhesive surface with ultraviolet rays or radiation, therebyintroducing a functional group having a cell adhesivity to theirradiated portion of the cell non-adhesive surface.

In still another aspect of the present invention, the cell arrangementcontrol device is prepared by patternwise irradiating a cell adhesive ornon-adhesive surface with ultraviolet rays or radiation, therebyintroducing polymerization initiation sites to the irradiated portion,and graft-polymerizing a monomer or monomers capable of providing apolymer having a cell adhesivity or a cell non-adhesivity onto theirradiated portion, thereby forming a pattern of the polymer.

It is possible to provide a fine pattern of cells in high resolution.Also since, according to the above processes, the patterned upper layerof the device is firmly fixed to the surface of the under layer bychemical bonding or a latent pattern is formed in a cell non-adhesivesurface, the patterned layer does not peel off during rinsing or cultureof cells and the accuracy of the pattern is maintained.

Controlled cell arrangement, namely a patterned cell layer, is easilyobtained by culturing cells on the thus obtained cell arrangementcontrol devices.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a microphotograph showing a pattern of stained cells;

FIG. 2 is a further enlarged microphotograph of the pattern shown inFIG. 1; and

FIG. 3 is an enlarged photograph of a photomask in the same enlargementratio as that of the photograph of FIG. 1.

DETAILED DESCRIPTION

In the cell arrangement control device of the present invention, apattern composed of a cell adhesive surface and a cell non-adhesivesurface is formed in or on the surface of a basis material constitutingthe device of the present invention (the device-constituting materialbeing hereinafter referred to as "substrate").

Examples of the material for the substrate are, for instance, variousplastics, glass, metals and ceramics, which may be in any form such asfilm, sheet, plate, dish, tube and other desired forms. Articles usedalready as devices or for other purposes, e.g. incubation dish andsemiconductor plate or board, can also be used in the present invention.

The cell adhesive surface includes, for instance, a surface having acharged functional group, e.g. carboxyl group or amino group, and/or acell adhesive peptide, e.g. RGDS (Arg-Gly-Asp-Ser), and a surface towhich a high molecular weight compound having a cell adhesivity(hereinafter referred to as "cell adhesive polymer") is fixed.

Examples of the cell adhesive polymer which provides the cell adhesivesurface are, for instance, a synthetic polymer having electric chargesuch as polyacrylic acid, polyvinyl sulfuric acid, polystyrene sulfonicacid or polyallylamine; a polysaccharide having electric charge such aschondroitin sulfate, dermatan sulfate, dextran sulfate, keratan sulfate,heparane sulfate, hyaluronic acid or chitin; a cell adhesive proteinsuch as collagen, gelatin, fibronectin or hydronectin; a cell adhesiveprotein-fixed synthetic polymer, a cell adhesive peptide-fixed syntheticpolymer, and other materials having a cell adhesivity. The cell adhesivepolymers may be used alone or in admixture thereof.

The surface having the functional group and/or cell adhesive peptide isformed in the surface of the substrate or on the surface of thesubstrate. That is to say, the functional group and/or peptide may beintroduced directly into the surface of the substrate or may beintroduced by fixing a high molecular weight compound having functionalgroup or peptide onto the surface of the substrate.

The direct introduction of the functional group such as carboxyl groupor amino group can be conducted in a known manner, for instance, byexposing the surface of the substrate made of plastics in a desired formsuch as dish, film or tube to ultraviolet rays in air or to radiationsuch as plasma, electron beams or γ-rays.

The fixing of the high molecular weight compound having functional groupand/or peptide to the substrate can be made by various processes, forinstance, a process wherein the functional group of the substrate andthe functional group of the high molecular weight compound are combinedby chemical reaction, a process wherein a monomer having a functionalgroup with electric charge such as carboxyl group is graft-polymerizedonto the surface of the substrate, a process wherein a monomer isgraft-polymerized onto the surface of the substrate, and carboxyl groupor amino group is introduced to the graft-polymerized polymer, forinstance, by graft-polymerizing acrylamide onto the substrate surface,and hydrolyzing the amide to carboxyl group by means of alkali treatmentor converting the amide to amino group by means of treating with analkali solution of sodium hypochlorite (Hofmann degradation).

The cell non-adhesive surface includes a hydrophobic surface having acontact angle of at least 90°, and a hydrophilic surface having noelectric charge and having a contact angle of at most 50°.

Examples of the hydrophobic surface are, for instance, surfaces ofpolytetrafluoroethylene and silicone.

Representative materials which provide the hydrophilic surface having acontact angle of at most 50° are hydrophilic polymers having no electriccharge. Examples of the hydrophilic polymer are, for instance, polyvinylalcohol, polyethylene glycol, polyacrylamide, polydimethylacrylamide,polyhydroxyethyl methacrylate, copolymers prepared from monomersconstituting the preceding hydrophilic polymers, and cellulose.

The hydrophobic surface and hydrophilic surface applicable to thepresent invention are not limited to those exemplified above.

The cell non-adhesive surface can be provided to the substrate, forinstance, by coating, laminating, photochemical fixing and otherappropriate processes. It is also possible to provide the cellnon-adhesive surface to the substrate by graft-polymerizing a monomerwhich is hydrophilic and has no electric charge to produce the cellnon-adhesive polymers as mentioned above.

When the substrate itself is made of the above-mentioned hydrophobic orhydrophilic material, it is not necessary to provide the cellnon-adhesive surface to the substrate.

The cell arrangement control device of the present invention can beprepared by processes utilizing a photosensitive hydrophilic polymer orby processes wherein patternwise irradiation of a cell adhesive ornon-adhesive surface is conducted in order to produce functional groupsor radicals.

In the first embodiment of the present invention, the cell arrangementcontrol device is prepared by a process which comprises (1) forming alayer of a photosensitive polymer having a cell non-adhesivity on a celladhesive surface by coating or causing to adsorb the photosensitivehydrophilic polymer, (2) placing a photomask having a desired pattern onthe photosensitive cell non-adhesive polymer layer and exposing to alight, and (3) developing the pattern by rinsing the photosensitive cellnon-adhesive polymer layer to form a pattern of the cell non-adhesivepolymer on the cell adhesive surface.

The photosensitive polymer having cell non-adhesivity used in thepresent invention can be a cell non-adhesive hydrophilic polymer towhich a photosensitive group is directly introduced, or a photosensitivecomposition containing a cell non-adhesive hydrophilic polymer and aphotosensitive compound. For instance, in the above process, the cellnon-adhesive hydrophilic polymer, preferably the above-mentionedhydrophilic polymer having no electric charge, can be easily fixedpatternwise on the cell adhesive surface, for instance, by forming alayer of a composition containing the cell non-adhesive hydrophilicpolymer and a compound having at least two azido groups on the celladhesive surface formed in or on the substrate, and patternwiseirradiating a light. Cell non-adhesive hydrophilic polymers to which atleast one azido group is directly introduced can be used, but the use ofthe composition containing the hydrophilic polymer and the azidocompound is preferred, since no special procedure for introducing azidogroups to the polymer is required and the removal of non-reactedsubstances is easy upon the development. It is preferable to use theazido compound in an amount of at least 1% by weight based on thepolymer.

As the compound having at least two azido groups, there can be used, forinstance, usual bisazido compounds as shown in Table 1 andazido-containing polymers wherein at least two azido groups areintroduced into one molecule of the polymers. However, the compoundhaving at least two azido groups used in the present invention is notlimited to them. The azido group includes, for instance, carbonylazidogroup (--CON₃), sulfonylazido group (--SO₂ N₃) and the aromatic azidogroup ##STR1## Above all, the aromatic azido group and sulfonylazidogroup are preferable because of the good stability. Particularly, acompound having aromatic azido group having an electron attractivesubstituent such as nitro group and a bisazido compound photosensitiveagainst i-line or g-line are preferable in respect that they can beconverted into nitrene by irradiation with a light having a relativelylonger wave length (>320 nm).

                                      TABLE 1                                     __________________________________________________________________________                                           Photosensible                          Bisazido compound                      range                                  __________________________________________________________________________     ##STR2##                              deep UV                                 ##STR3##                              "                                       ##STR4##                              "                                       ##STR5##                              "                                       ##STR6##                              "                                       ##STR7##                              "                                       ##STR8##                              "                                       ##STR9##                              i-line                                  ##STR10##                             i-line                                  ##STR11##                             "                                       ##STR12##                             "                                       ##STR13##                             "                                       ##STR14##                             "                                       ##STR15##                             "                                       ##STR16##                             g-line                                 __________________________________________________________________________

The cell non-adhesive hydrophilic polymer is fixed to the cell adhesivesurface through the azido compound by a reaction wherein the azidogroups are converted into nitrene groups by light irradiation and thenitrene groups undergo chemical reactions as shown by the followingreaction formulas with the cell adhesive surface and the hydrophilicpolymer, namely a hydrogen-abstraction reaction represented by theformula (1), an insertion into C--H bond or an addition to double bondrepresented by the formula (2) and coupling reaction represented by theformula (3). ##STR17##

The nitrene group is so highly reactive that bonding may be formedthrough other reactions than those above-mentioned. Also theabove-mentioned reactions may occur between the hydrophilic polymermolecules to cause crosslinking. There is a case where the cellnon-adhesive polymer is more stably fixed to the cell adhesive surfaceby occurrence of crosslinking.

It is not always required that the cell non-adhesive hydrophilic polymeris fixed to the cell adhesive surface by chemical bonding as mentionedabove, but it may be adhered as a film onto the cell adhesive surface.

The application of the photosensitive composition to the cell adhesivesurface can be made, for instance, by a method wherein thephotosensitive composition is dissolved or dispersed in a volatileorganic solvent such as methanol, the solution or dispersion is coatedby spreading or spraying onto the cell adhesive surface and it is driedto form a thin layer of the composition on the cell adhesive surface, ora method wherein the cell adhesive surface is brought into contact withan aqueous solution or colloidal solution of the photosensitivecomposition so as to adsorb the composition, for instance, by immersionin the solution of by causing the solution to flow on the cell adhesivesurface. The film formation by casting from the solution in a volatileorganic solvent such as methanol is preferred because a uniform thinlayer is easily obtained.

Also, there may be usable a method wherein the compound having at leasttwo azido groups is applied onto the cell adhesive surface and the cellnon-adhesive hydrophilic polymer is then applied.

As a light source for the irradiation operation, there can be usedvarious kinds of mercury light such as a high pressure mercury lamp, alow pressure mercury lamp, an extra-high pressure mercury lamp; excimerlaser; and the like. In case of using azido compounds sensitive to thelonger wavelength region in the ultraviolet region, damages by theshorter wavelength ultraviolet rays to the hydrophilic polymer, the celladhesive surface or the device material can be reduced by cutting offthe shorter wavelength region using a filter. This method isparticularly preferable in case that hydrophilic polymers such asprotein are used.

The reaction of a nitrene group is accomplished in a very short time,and irradiation for five minutes is enough for the reaction.

The patternwise irradiation can be made by various methods, forinstance, a method wherein a light is irradiated through a photomaskhaving a pattern placed on the photosensitive layer, or a methodutilizing lithography by means of excimer laser.

In the second embodiment of the present invention, the cell arrangementcontrol device is prepared in the same manner as above except thatinstead of forming the layer of the photosensitive cell non-adhesivepolymer on the cell adhesive layer in the step (1), a layer of aphotosensitive polymer having cell adhesivity is formed on a cellnon-adhesive surface by coating or causing to adsorb the photosensitivepolymer. As the photosensitive polymer, there can be used the celladhesive polymers mentioned above to which at least one azido group isdirectly introduced or a photosensitive composition containing theabove-mentioned cell adhesive polymers and the above-mentioned azidocompounds having at least two azido groups. For instance, the celladhesive hydrophilic polymer can be easily fixed patternwise onto thecell non-adhesive surface by applying the photosensitive composition tothe cell non-adhesive surface formed on the substrate, and patternwiseirradiating a light such as ultraviolet rays.

According to the above-mentioned first and second processes of thepresent invention, the patterned upper layer is covalently bondedthrough nitrogen atom derived from azido group to the cell adhesive ornon-adhesive surface of the under layer, thus the pattern is firmlyfixed to the under layer.

The cell arrangement control device can also be prepared by a processutilizing irradiation of ultraviolet rays or radiation wherein afunctional group which renders a surface cell-adhesive, or a sitecapable of initiating a polymerization is formed in a cell adhesive ornon-adhesive surface by patternwise irradiation of ultraviolet rays orradiation.

In the third embodiment of the present invention, the device is preparedby a process which comprises placing a mask having a desired pattern onthe cell non-adhesive surface of the substrate, and irradiatingultraviolet rays or radiation to the cell non-adhesive surface throughthe mask, thereby producing a cell adhesive functional group only in theexposed portion of the cell non-adhesive surface to patternwise form acell adhesive surface in the cell non-adhesive surface.

The cell adhesive functional groups formed in this process topatternwise render the cell non-adhesive surface cell-adhesive arecarboxyl group and amino group. As the cell non-adhesive surface used inthis process, there are used hydrophilic polymers having no electriccharge and capable of producing carboxyl group or amino group, e.g.polydimethylacrylamide, among the hydrophilic polymers exemplifiedbefore.

The formation of the functional group can be conducted in a knownmanner, for instance, by irradiation of ultraviolet rays in the air ortreatment with radiation. Examples of the radiation are, for instance,plasma obtained by corona discharge or glow discharge, electron beams,γ-rays, and other high energy rays. The use of the plasma obtained byglow discharge is preferred in view of its applicability to variousmaterials and the effect, and N₂ plasma and O₂ plasma are particularlypreferred in view of efficiency in producing the functional groups. Theuse of ultraviolet rays is also preferable in that the treatment in highdegree of resolution is possible.

The masks used in the invention are those having an openingcorresponding to a pattern to be delineated and capable of causing onlythe portion corresponding to the opening to expose to ultraviolet raysor radiation irradiated, and general photomasks can be utilized.Particularly, in case of using radiation such as plasma, a mask made ofa non-metallic material is preferable, since the use of a metallic maskmay be inferior in degree of resolution owing to discharge from themask.

In the fourth embodiment of the present invention, the cell arrangementcontrol device is prepared by a process which comprises placing a maskhaving a desired pattern on a cell adhesive or non-adhesive surface ofthe substrate, irradiating ultraviolet rays or radiation to the surfacethrough the mask, thereby producing a polymerization initiation siteonly in the exposed portion, and graft-polymerizing a cell adhesive ornon-adhesive monomer on the treated surface, thereby producing thepolymer only on the exposed portion in the form of the pattern.

The polymerization initiation site induced by the irradiation in thisprocess is a polymer radical derived from the material of the substrateitself or the polymer layer provided on the substrate. The polymerradical can be induced in a known manner by treatment with ultravioletrays or radiation such as plasma. Since UV treatment generally requiresthe use of sensitizers, the radiation treatment which does not requirethe use of sensitizers is preferred.

The radiations mentioned above can be used in this process. Above all,plasma as obtained by glow discharge is preferable.

The graft polymerization of cell adhesive monomers or cell non-adhesivemonomers based on the thus induced radicals can be conducted, forinstance, by directly introducing the monomers into the system where theradicals are produced, or by once bringing the radicals into contactwith air to convert into peroxide, adding the monomers to apolymerization system, and decomposing the peroxide into radicals toinitiate the polymerization.

Examples of the cell adhesive monomer are, for instance, monomers havingelectric charge such as acrylic acid, styrene sulfonic acid and vinylsulfonic acid. Examples of the cell non-adhesive monomer are, forinstance, hydrophilic monomers having no electric charge such asacrylamide, dimethylacrylamide, ethylene glycol and hydroxyethylmethacrylate.

The photomasks mentioned above are also applicable to this process.

The arrangement of cells can be easily controlled by a cell culture in ausual manner using the cell arrangement control devices prepared asmentioned above. It is possible to form a fine pattern of cells of ahigh resolution on the order of up to μm.

The obtained fine patterns are useful for the production of ultra-smallbiosensor, switching device, bioreactor, hybrid type artificial organand so on, and the development of neuro-computer. The devices are alsoapplicable to research of cell functions such as informationtransmission between cells.

The present invention is more specifically described and explained bymeans of the following examples, in which all parts and % are by weightunless otherwise noted. It is to be understood that the presentinvention is not limited to these examples.

EXAMPLE 1

N,N-Dimethylacrylamide was polymerized in acetone in the presence of aredox catalyst system of benzoyl peroxide and N,N-dimethyl-p-toluidineto produce poly(N,N-dimethylacrylamide)(hereinafter referred to as"PDMAA").

A mixture of 95 parts of PDMAA and 5 parts of sodium4,4'-diazidostilbene-2,2'-disulfonate as the bisazido compound wasdissolved in methanol to form a 0.1% solution.

The obtained solution was cast onto a polystyrene dish for tissueculture and dried to form a film having a thickness of several tens ofnm. On the thus formed film was placed a photomask as shown in FIG. 3having slits wherein the width of a pair of the opening portion and thenon-opening portion was 250 μm, and the film was exposed through thephotomask to a light from a high pressure mercury lamp (400 W) for 30seconds (corresponding to light quality of about 600 mJ/cm² measured bya detector for i-line 365 nm).

The exposed film was then developed by thoroughly rinsing with methanoland water to remove the non-exposed portion of PDMAA film, thus the dishhaving a fine pattern composed of PDMAA and the dish surface.

By using DMEM (Dulbecco's Modified Eagle's Medium) containing fetal calfserum as a culture medium, bovine vascular endothelial cells wereinoculated into the thus obtained dish, and cultured in a CO₂ incubatorat 37° C. The endothelial cells selectively spread and grew only on thePDMAA-removed portion (non-exposed portion), thus the cell pattern shownin FIGS. 1 and 2 was obtained, wherein FIG. 1 was a photograph of thepattern of the stained cells (enlargement ratio being the same as FIG.3) and FIG. 2 was a photograph of further enlarged pattern shown in FIG.1.

EXAMPLE 2

A 0.1% methanol solution of PDMAA and the bisazido compound prepared inthe same manner was cast on a polystyrene dish and dried to form a thinfilm. The PDMAA film was irradiated with ultraviolet rays from a highpressure mercury lamp to fix the film onto the surface of the dish.

A copolymer of 80% of N,N-dimethylacrylamide and 20% ofacryloyloxysuccinimide was reacted with fibronectin in a phosphatebuffer (pH 8.5) to produce a fibronectin-fixed N,N-dimethylacrylamidecopolymer (hereinafter referred to as "FN-PDMAA").

A mixture of 95 parts of FN-PDMAA and 5 parts of sodium4,4'-diazidostilben-2,2'-disulfonate was dissolved in methanol to give a0.1% solution. The resulting solution was cast on the PDMAA-fixed dishand dried to form a FN-PDMAA film having a thickness of several tens ofnm. The FN-PDMAA film containing bisazido compound was then exposedthrough a photomask to ultraviolet rays from a high pressure mercurylamp for 30 seconds.

The exposed film was then developed by thoroughly rinsing with methanoland water to remove the masked portion of the FN-PDMAA film, thusobtaining a dish having a fine pattern composed of the FN-PDMAA and thePDMAA surface.

Bovine vascular endothelial cells were cultured in the same manner as inExample 1 by using the thus obtained dish. The cells selectively spreadand grew only on the FN-PDMAA fixed portion (exposed portion) to give acell pattern.

EXAMPLE 3

A 0.1% methanol solution of 95 parts of PDMAA and 5 parts of sodium4,4'-diazidostilbene-2,2'-disulfonate was cast on a polystyrene Schaleand dried to give a PDMAA film containing the bisazido compound. ThePDMAA film was then irradiated with ultraviolet rays from a highpressure mercury lamp to chemically fix PDMAA onto the surface of theSchale.

The same photomask as used in Example 1 was placed on the PDMAA film.The PDMAA film covered with the photomask was subjected to glowdischarge treatment in an ion coater (IB-3 type ion coater made by EikoEngineering Co., Ltd.) (0.2 Torr, 5 mA×10 seconds), and was then allowedto stand in contact with air to produce carboxyl group in the exposedportion of the PDMAA film.

Bovine vascular endothelial cells were cultured on the thus obtainedSchale in the same manner as in Example 1. The cells selectively grewonly on the carboxyl group introduced portion (exposed portion).

EXAMPLE 4

A PDMAA-fixed Schale prepared in the same manner as in Example 3 wasirradiated in air with ultraviolet rays from a low pressure mercury lampfor 1 hour through the photomask shown in FIG. 3 to induce carboxylgroup in the exposed portion of the PDMAA film.

Bovine vascular endothelial cells were cultured on the thus treatedSchale in the same manner as in Example 1. The cells selectively grewonly on the carboxyl group-introduced portion (exposed portion) to givea cell pattern.

EXAMPLE 5

A polystyrene Schale for tissue culture use was covered with thephotomask shown in FIG. 3, subjected to the glow discharge treatment inan ion coater and then brought into contact with air in order tointroduce peroxide into the exposed portion of the polystyrene Schale.

The Schale was then immersed in a 5% aqueous solution of acrylamide, andafter thoroughly degasing, the system was sealed and the polymerizationof acrylamide was conducted at 60° C. for 2 hours. After the completionof the polymerization, the Schale was washed well with water to give atissue culture polystyrene Schale having polyacrylamide grafted onlyonto the exposed portion of the Schale.

The culture of cells was carried out on the thus obtained Schale in thesame manner as in Example 1. The cells selectively grew only on thenon-grafted portion (non-exposed portion), thus the same cell pattern asthat shown in FIGS. 1 and 2 was obtained.

EXAMPLE 6

The mask shown in FIG. 3 was placed on a polytetrafluoroethylene film,and the film was subjected to glow discharge treatment in the samemanner as in Example 3 to introduce peroxide into the exposed portion ofthe film. Acrylamide was then graft-polymerized only onto the exposedportion in the same manner as in Example 5.

The thus obtained polyacrylamide-grafted polytetrafluoroethylene filmwas immersed in a 1N-NaOH aqueous solution at 60° C. for 1 hour tohydrolyze the amido into carboxyl group.

Bovine vascular endothelial cells were cultured on the obtained film inthe same manner as in Example 1. The cells selectively grew only on thepolymer-grafted portion (exposed portion) to give a cell pattern.

EXAMPLE 7

A polyacrylamide-grafted polytetrafluoroethylene film was prepared inthe same manner as in Example 6, and it was immersed in an aqueous NaOHalkaline solution of sodium hypochlorite and kept at 0° C. for 1 hoursto convert the amido portion into amino group by means of Hofmanndegradation.

Bovine vascular endothelial cells were cultured on the thus obtainedfilm in the same manner as in Example 1. The cells selectively grew onlyin the polymer-grafted portion (exposed portion) to give a cell pattern.

In addition to the ingredients used in the examples, other ingredientscan be used in the examples as set forth in the specification to obtainsubstantially the same results.

What we claim is:
 1. A process for preparing a cell arrangement control device which consists essentially of applying a photosensitive, cell non-adhesive polymer to a cell adhesive surface, irradiating patternwise the resulting photosensitive cell non-adhesive polymer layer with a light, and developing the irradiated layer to leave the irradiated portion on said cell adhesive surface, thereby providing a pattern of said cell non-adhesive polymer on said cell adhesive surface.
 2. The process of claim 1, wherein said photosensitive polymer is a cell non-adhesive polymer having at least one azido group.
 3. The process of claim 1, wherein said photosensitive polymer is a composition containing a cell non-adhesive polymer and a compound having at least two azido groups. 